title: Gene delivery with active targeting to ovarian cancer cells mediated by folate receptor alpha.
abstract: Folate receptor alpha (FRalpha) is overexpressed on ovarian cancer cells and is a promising molecular target for ovarian cancer gene therapy, but there was still no related report. In this study, folate modified cationic liposomes (F-PEG-CLPs) for ovarian cancer gene delivery were developed for the first time. Folate-poly(ethylene glycol)-succinate-cholesterol (F-PEG-suc-Chol) was firstly synthesized and then used to prepare folate-targeted cationic liposomes/plasmid DNA complexes (F-targeted lipoplexes). F-targeted lipoplexes were prepared by post-insertion method, and displayed membrane structure by transmission electron microscopy observation with the diameter of 193 nm-200 nm and the zeta potential of 35 mV-38 mV. DNase degradation experiments showed that plasmid DNA could be effectively shielded by F-targeted lipoplexes in vitro. F-targeted lipoplexes could transfer gene into human ovarian carcinoma cell line SKOV-3, and 0.1% F-PEG-CLPs composed by DOTAP/Chol/mPEG-Chol/F-PEG-suc-Chol (50:45:5:0.1, molar ratio) had the highest transfection efficiency. The transfection activity of F-targeted lipoplexes could be competitively inhibited by free folic acid, demonstrating that folate-FRalpha interaction caused high transfection efficiency of F-targeted lipoplexes. The uptake mechanism of F-targeted lipoplexes was further validated on human oral carcinoma cell line KB and human liver carcinoma cell line HepG2. The concentration-dependent and time-dependent cytotoxicity of targeted material F-PEG-suc-Chol was observed by MTT assay on SKOV-3 cell and its application would not increase the cytotoxicity of F-targeted lipoplexes in SKOV-3 cells. All the data indicated that F-PEG-CLPs would be a promising gene vector targeting for ovarian cancer therapy.
authors.affiliation: State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China
authors.forename: Zhiyao
authors.initials: Z
authors.lastname: He
authors.forename: Yiyi
authors.initials: Y
authors.lastname: Yu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yongdong
authors.initials: Y
authors.lastname: Yan
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
authors.forename: Jun
authors.initials: J
authors.lastname: He
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Xie
authors.forename: Gu
authors.initials: G
authors.lastname: He
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
authors.forename: Xiangrong
authors.initials: X
authors.lastname: Song
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology; D002784:Cholesterol / Q000008:administration & dosage / Q000494:pharmacology; D005260:Female; D058975:Folate Receptor 1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D005492:Folic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D007624:KB Cells; D008081:Liposomes; D008954:Models, Biological; D058990:Molecular Targeted Therapy; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D011092:Polyethylene Glycols / Q000008:administration & dosage* / Q000494:pharmacology; D019802:Succinic Acid / Q000008:administration & dosage / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; C548696:FOLR1 protein, human; D058975:Folate Receptor 1; D008081:Liposomes; C552831:poly(ethylene glycol)-folate; D011092:Polyethylene Glycols; D005492:Folic Acid; D002784:Cholesterol; D019802:Succinic Acid